Differentiation of human pluripotent stem cells (hPSCs) requires
precise signaling control at each step to mimic the embryonic
development. Advances in using highly specific and potent small molecule
modulators of signaling pathways have allowed efficient generation of
nearly homogenous populations of specific cell types from hPSCs. For
highly efficient conversion of hESCs/iPSCs into neural precursor cells
in a monolayer fashion, inhibition of both BMP and TGFβ signaling
pathways (i.e., dual SMAD inhibition) by small molecule inhibitors, LDN-193189 (0.1 µM) and SB431542
(10 µM) respectively (i.e., Neural-LSB), was developed as the most
effective condition. Under this dual SMAD inhibition using the
Neural-LSB small molecule cocktail, more than 80% of hPSCs are rapidly
induced into Pax6+ neural precursor cells. This condition can be further
combined with other patterning cues to generate subtype specific neural
and neuronal cells, such as floor plate and midbrain dopaminergic
neurons using Hedgehog and Wnt pathway activators (i.e., purmorphamine
and CHIR99021), or nociceptors using 3i (combination of CHIR99021/GSK3
inhibitor, SU5402/FGF-VEGF-PDGF inhibitor, DAPT/Notch inhibitor).

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Differentiation of human pluripotent stem cells (hPSCs) requires
precise signaling control at each step to mimic the embryonic
development. Advances in using highly specific and potent small molecule
modulators of signaling pathways have allowed efficient generation of
nearly homogenous populations of specific cell types from hPSCs. For
highly efficient conversion of hESCs/iPSCs into neural precursor cells
in a monolayer fashion, inhibition of both BMP and TGFβ signaling
pathways (i.e., dual SMAD inhibition) by small molecule inhibitors, LDN-193189 (0.1 µM) and SB431542
(10 µM) respectively (i.e., Neural-LSB), was developed as the most
effective condition. Under this dual SMAD inhibition using the
Neural-LSB small molecule cocktail, more than 80% of hPSCs are rapidly
induced into Pax6+ neural precursor cells. This condition can be further
combined with other patterning cues to generate subtype specific neural
and neuronal cells, such as floor plate and midbrain dopaminergic
neurons using Hedgehog and Wnt pathway activators (i.e., purmorphamine
and CHIR99021), or nociceptors using 3i (combination of CHIR99021/GSK3
inhibitor, SU5402/FGF-VEGF-PDGF inhibitor, DAPT/Notch inhibitor).